SMC1 Overexpression Lysate Summary
Description |
SMC1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC215888
Accession#: NM_006306
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
SMC1A |
Applications/Dilutions
Dilutions |
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Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Theoretical MW |
143.1 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for SMC1 Overexpression Lysate
Background
Proper cohesion of sister chromatids is a prerequisite for the correct segregation of chromosomes during cell division. The cohesin multiprotein complex is required for sister chromatid cohesion. This complex is composed partly of two structural maintenance of chromosomes (SMC) proteins, SMC3 and either SMC1L2 or the protein encoded by this gene. Most of the cohesin complexes dissociate from the chromosomes before mitosis, although those complexes at the kinetochore remain. Therefore, the encoded protein is thought to be an important part of functional kinetochores. In addition, this protein interacts with BRCA1 and is phosphorylated by ATM, indicating a potential role for this protein in DNA repair. This gene, which belongs to the SMC gene family, is located in an area of the X-chromosome that escapes X inactivation. [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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Publications for SMC1 Lysate (NBL1-16236) (0)
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Product General Protocols
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Video Protocols
FAQs for SMC1 Lysate (NBL1-16236). (Showing 1 - 2 of 2 FAQ).
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I would like to use one of your anti-smc1 antibodies to detect smc1 in an invertebrate. Your different antibodies are raised against different peptides derived from different regions possibly well-conserved, but not completely, in the organism that interests me (Schistosoma mansoni). Would it be possible to know the sequence of the different peptides that was used to generate the different antibodies, or failing that, could you analyze the corresponding sequence in my species of interest in order to determine whether or not it corresponds to one of the different peptides?
- My suggestion to you would be to look at the homology between the protein expressed in your species and the protein expressed in the reactive species of the antibodies. A homology of >90% is a great start, >80% is acceptable. Please view our list of smc1 antibodies we currently offer and there are 27 products.
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My sequence of interest have >80% homology with the N-term and the C-term parts of human smc1. Your antibodies NB100-78322 and NB100-204 recognized the N-term and C-term of human smc1 respectively, therefore it’s possible that the cross reactivity can happen. I would be very interested to test these antibodies (NB100-78322 and NB100-204). Can you provide me free sample (few ul) to do western blot. I will share the testing result with you.
- We do not provide free samples for testing. If you are interested in testing these antibodies in an unlisted species or application, you would qualify for the Innovator's Rewards program. The way the program works is that you would purchase the antibody of interest, perform your testing using appropriate controls and then share your data with us. Upon review of your data we would issue you a voucher for a free antibody. You can find more details here.
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